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Updated: Sep 20, 2025
In Vivo CRISPR/Cas9 Screening to Simultaneously Evaluate Gene Function in Mouse Skin and Oral Cavity
Published on: November 2, 2020
In Vivo CRISPR Interference Screen Reveals Long Noncoding RNA Portfolio Crucial for Cutaneous Squamous Cell Carcinoma
Gyuhyeon Kim1, Zurab Siprashvili2, Xue Yang2
1Program in Epithelial Biology, Department of Dermatology, Stanford University School of Medicine, Stanford, California, USA; Department of Biochemistry, Stanford University School of Medicine, Stanford, California, USA.
Abstract:
Cutaneous squamous cell carcinoma (cSCC) accounts for 20% of all skin cancer mortality globally, making it the second-highest subtype of skin cancer. The high prevalence of cSCC in humans highlights the need to uncover alternative actors and mechanisms influencing skin cancer development. Significant advances have been made to better understand some key factors in cSCC growth. However, little is known about the role of noncoding RNAs, particularly of a specific subclass termed long noncoding RNA (lncRNA). By performing pseudobulk analysis of single-cell sequencing data from normal and cSCC human skin tissues, we determined a global portfolio of lncRNAs specifically expressed in keratinocyte subpopulations. Integration of CRISPR interference screens in vitro and the xenograft model identified several lncRNAs impacting the growth of cSCC cancer lines both in vitro and in vivo. Among these, we further validated LINC00704 and LINC01116 as proliferation-regulating lncRNAs in cSCC lines and potential biomarkers of cSCC growth. Taken together, our study provides a comprehensive signature of lncRNAs with roles in regulating cSCC growth.
Insights
This study identifies long noncoding RNAs (lncRNAs) that regulate the growth of cutaneous squamous cell carcinoma (cSCC). LINC00704 and LINC01116 are highlighted as key players and potential biomarkers for this common skin cancer.
Area of Science:
- Oncology
- Molecular Biology
- Dermatology
Background:
- Cutaneous squamous cell carcinoma (cSCC) is a significant cause of skin cancer mortality.
- Understanding novel molecular mechanisms driving cSCC growth is crucial.
- The role of long noncoding RNAs (lncRNAs) in cSCC remains largely unexplored.
Purpose of the Study:
- To identify lncRNAs involved in regulating cSCC development.
- To characterize the function of specific lncRNAs in cSCC cell proliferation.
- To explore potential lncRNA biomarkers for cSCC.
Main Methods:
- Pseudobulk analysis of single-cell sequencing data from normal and cSCC human skin.
- CRISPR interference screens in vitro.
- Xenograft models for in vivo validation.
Main Results:
- A global portfolio of keratinocyte-specific lncRNAs in cSCC was determined.
- Several lncRNAs were identified that impact cSCC growth in vitro and in vivo.
- LINC00704 and LINC01116 were validated as proliferation-regulating lncRNAs in cSCC.
Conclusions:
- This study provides a comprehensive signature of lncRNAs implicated in cSCC growth regulation.
- LINC00704 and LINC01116 show potential as biomarkers for cSCC.
- lncRNAs represent a novel class of therapeutic targets for cSCC.
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